College of Science and Technology, Nottingham Trent University, Nottingham, UK.
Department of Biochemistry and Genetics, University of Navarra, 31008, Pamplona, Spain.
Sci Rep. 2019 Jan 23;9(1):390. doi: 10.1038/s41598-018-36888-x.
AGAP2 (Arf GAP with GTP-binding protein-like domain, Ankyrin repeat and PH domain 2) isoform 2 is considered a proto-oncogene, but not much is known about AGAP2 gene expression regulation. To get some insight into this process, AGAP2 proximal promoter was cloned and characterised using reporter assays. We have identified SP1 as a transcription factor bound to AGAP2 promoter and required for AGAP2 expression in two different types of cancer cells (KU812, a chronic myeloid leukaemia cell line; and DU145, a prostate cancer cell line): silencing SP1 decreased AGAP2 protein levels. We have also found that all-trans retinoic acid (ATRA) treatment increased AGAP2 protein levels in both cell lines whilst curcumin treatment reduced ATRA-mediated AGAP2 increase. Furthermore, chromatin immunoprecipitation studies revealed the presence of RARα, RXRα and the lysine acetyl transferase PCAF in AGAP2 promoter. Our results provide a novel understanding of AGAP2 expression regulation that could be beneficial to those patients with cancers where AGAP2 is overexpressed.
AGAP2(富含 GTP 结合蛋白结构域、锚蛋白重复和 PH 结构域的 Arf GAP2 同工型 2)同工型 2 被认为是一种原癌基因,但对于 AGAP2 基因表达调控知之甚少。为了深入了解这一过程,我们使用报告基因检测克隆和鉴定了 AGAP2 近端启动子,并进行了特征分析。我们已经确定 SP1 是一种与 AGAP2 启动子结合的转录因子,对于两种不同类型的癌细胞(KU812,慢性髓性白血病细胞系;和 DU145,前列腺癌细胞系)中 AGAP2 的表达是必需的:沉默 SP1 会降低 AGAP2 蛋白水平。我们还发现,全反式维甲酸(ATRA)处理会增加两种细胞系中的 AGAP2 蛋白水平,而姜黄素处理会降低 ATRA 介导的 AGAP2 增加。此外,染色质免疫沉淀研究表明,RARα、RXRα 和赖氨酸乙酰转移酶 PCAF 存在于 AGAP2 启动子中。我们的研究结果提供了对 AGAP2 表达调控的新认识,这可能对那些 AGAP2 过表达的癌症患者有益。